Physical Sciences and Mathematics Commons^™

Characterization Of Small Molecules And Ions By Atmospheric Pressure Ionization-Mass Spectrometry, Maleesha De Silva

Theses and Dissertations

Mass spectrometry (MS) has become a widely used technique for the characterization of a wide range of substances in diverse fields. The selection of appropriate ionization techniques, source parameters, charge carriers based on the analyte’s polarity is essential in MS as only the ions are detected. When using a soft ionization technique, single stage-MS at best provides only chemical composition; thus, tandem MS is needed to determine structural information and dissociation pathways. This dissertation focuses on the characterization of various small molecules and ions using different ionization techniques, charge carriers, and collision-induced dissociation (CID).

In Chapter II, commercially available ionic …

Mass Spectrometry Directed Pre-Clinical Studies Of Asthma Candidate Midd0301 And Its Analogs, Md Shadiqur Rashid Roni

Theses and Dissertations

ABSTRACTMASS SPECTROMETRY DIRECTED PRE-CLINICAL STUDIES OF ASTHMA CANDIDATE MIDD0301 AND ITS ANALOGS by Md Shadiqur Rashid Roni The University of Wisconsin-Milwaukee, 2021 Under the Supervision of Professor Alexander (Leggy) Arnold

Asthma is one of the most prevalent global health problems. More than 25 million Americans are suffering from asthma and their symptoms especially in children are not well controlled with current medications. The current therapeutic options to control symptoms are limited and are commonly based on inhaled corticosteroids. There are several reasons for uncontrolled asthma symptoms that includes steroid resistance, improper use of inhalers and very few orally available medications. …

Mechanisms And Applications Of Improved Protein Analysis By Desorption Electrospray Ionization Mass Spectrometry (Desi-Ms), Roshan Javanshad

Dissertations

Electrospray ionization mass spectrometry (ESI-MS) is a soft ionization technique that allows detection of macromolecules, such as intact proteins, by the formation of multiply charged ions from solutions. Desorption electrospray ionization mass spectrometry (DESI-MS) is an ambient ionization technique that directly samples analyte from a surface during ESI-MS analysis. Although DESI-MS is highly accomplished at the analyses of metabolites, lipids, and other small molecules, it is far more limited when it comes to protein analysis. While most of the field in ambient ionization MS has moved towards primarily applications, our approach has been to explore the use of DESI-MS and …

Characterization Of Biological Samples Using Multi-Modal Mass Spectrometry, Yarixa L. Cintron-Diaz

FIU Electronic Theses and Dissertations

There is a need for fast, accurate and cost-effective protocols capable of assessing biomolecules at the molecular level (e.g., proteins, lipids and metabolites) from biological specimens. Mass spectrometry (MS) based techniques have become the analytical gold standard for identification and characterization of biomarkers in biological samples. The high throughput and short analysis time scales enables to follow biological processes while providing detailed chemical and spatial characterization. One of the current challenges in biological MS, is the high molecular complexity, chemical diversity and dynamic range. In this dissertation, the use of multi-modal mass spectrometry workflows -mass spectrometry imaging and ion mobility …

Non-Target Analysis Using High-Resolution Mass Spectrometry To Characterize And Remediate Urban Waters, Brian Ng

FIU Electronic Theses and Dissertations

The first part of this dissertation will focus on the development of a simple, robust online solid phase extraction liquid chromatography high resolution mass spectrometry (SPE LC-HRMS) method followed by the use of computational software workflows for non-target analysis (NTA) of environmental samples (development). The benchmarks to assess reproducibility are not well defined for non-target analysis. Parameters to evaluate analytical performance, such as accuracy, precision and selectivity, are well defined for target analysis, but remain elusive for non-target screening analysis. In this study, quality control (QC) guidelines are proposed to assure reliable data in NTA methodologies using a simple …

Investigating The Chemistry Of Harmful Algal Blooms And Microbial Communities In The Natural Environment, Samuel Patrick Putnam

Theses and Dissertations

Cyanobacteria has existed on the planet for over 3 billion years, and as technology, science, and industry have allowed us to shape our world, these changes have caused a proliferation of biomass. Higher carbon dioxide levels in the atmosphere, warmer temperatures, and rapid increases in the nutrient cycle have created excellent growing conditions for cyanobacteria, making them endemic in fresh waters throughout the world. The work in this dissertation was directed towards the secondary metabolites produced by cyanobacteria and other surrounding microbes and understanding their concentrations and release in natural systems. Analytical methods were developed for the extraction and quantification …

Don't Sell Them Short, There's More To Bacterial Natural Products Than Antibiotics, Alison Clare Domzalski

Dissertations, Theses, and Capstone Projects

Recent genomic studies of microbiomes have revealed an overwhelming number of biosynthetic genes of unknown function. Most of these “cryptic” biosynthetic genes are not expressed in laboratory monocultures of individual microbes. Thus, there remains tremendous untapped potential for natural products discovery. Here we employ mixed microbial culture (MMC) as a simple yet powerful approach to awaken cryptic biosynthetic gene clusters. Our preliminary studies demonstrated that arrays of metabolites could be induced in MMCs upon environmental cues, such as surface adhesion. Using this system, we have screened, identified, and isolated bioactive bacterial metabolites, which were characterized structurally and biologically. Of the …

Mass Spectrometry Based Metabolomics And Lipidomics., Fang Yuan

Electronic Theses and Dissertations

Metabolomics and lipidomics studies are important to understand living organisms at the molecular level. In this dissertation, I conducted various untargeted and targeted metabolomics and lipidomics studies using mass spectrometry based analytical platforms. Firstly, I investigated the androgen depletion effects on fecal metabolites in a mouse model of systemic lupus erythematosus and metabolite biomarkers for colorectal cancer using two-dimensional gas and liquid chromatography-mass spectrometry. Secondly, I investigated the relationship between gut microbiota-mediated resistance and susceptibility to malaria and the production of short-chain fatty acids using gas chromatography-mass spectrometry. Thirdly, I developed a high-throughput liquid chromatography-mass spectrometry method to profile long-chain …

Records Of Enriched Uranium Atmospheric Deposition In Pond Sediments In Piketon, Oh, Brianna Herner, Brian Majestic, Michael Ketterer

DU Undergraduate Research Journal Archive

The enrichment of uranium, often for nuclear weapons programs and commercial nuclear reactors, produces higher concentrations of radioactive uranium 235 (²³⁵U) than what naturally occurs, which can pose a human health hazard. The most abundant naturally occurring uranium isotope is ²³⁸U, which is still radioactive, however a higher concentration of ²³⁵U skews the observed isotopic uranium distribution. The Department of Energy Portsmouth Gaseous Diffusion Plant, located near Piketon, OH, enriched uranium from 1954 to 2001 and ²³⁵U pollution has recently been detected in air and sediment samples in the surrounding community. The extent of the …

Integrated Mass Spectrometry-Based Method In Protein Chemistry: Metal-Binding Protein And Integral Membrane Protein, Chunyang Guo

Arts & Sciences Electronic Theses and Dissertations

Most biological processes are associated with protein catalysis. Characterizing protein structure, therefore, is crucial for understanding biological function. Mass spectrometry (MS)-based methods have emerged as a pivotal biophysical tool to interrogate protein structures; they can characterize protein-ligand/protein complexes that are refractory to conventional high-resolution means such as X-ray crystallography, Cryo-EM, and nuclear magnetic resonance (NMR) spectroscopy, in part owing to high molecular weight and significant flexibility. The integrated MS-based platform will provide topology and structural information of proteins from sketch to detail; native MS identifies protein complexes’ composition and stoichiometry; hydrogen-deuterium exchange MS (HDX-MS) detects the protein complexes’ dynamic change …

Mass Spectrometry-Based Strategies In Protein Higher Order Structure Analysis: Fundamentals And Applications In Protein-Ligand Interactions, Xiaoran Liu

Arts & Sciences Electronic Theses and Dissertations

Protein ligand interaction is a fundamental question in biology and biochemistry, and many approaches including X-ray crystallography, nuclear magnetic resonance, cryogenic electron microscopy, mass spectroscopy (MS), infrared spectroscopy, circular dichroism, fluorescence spectroscopy and many others have been applied to address this question. Among these techniques, mass spectroscopy has the advantage of high throughput, low sample amount requirement, and mid-to-high spatial resolution. One of the MS-based approaches is protein footprinting, which utilizes labeling reagents to map the solvent accessible surface of the protein of interest thus deliver structural information. Irreversible labeling is represented by covalent labeling and radical labeling, in which …

Validation Of A Deployable Proteomic Assay For The Serological Screening Of Sexual Assault Samples, Catherine O'Sullivan Brown

Electronic Theses and Dissertations

Protein mass spectrometry (MS) has emerged as a technique to supplant traditional serological tests for body fluid identification. It was hypothesized that proteomic techniques would surpass the sensitivity and specificity of traditional serological techniques. An automated workflow coupled with protein MS has been developed for the confirmatory identification of five biological fluids. A developmental validation was completed, assessing parameters such as reproducibility, sensitivity, ion suppression, and limit of detection. Implementation was determined through tandem sample processing by MS, traditional serological tests, and standard DNA profiling methods. The MS approach offered superior detection limits while also providing true confirmatory results, producing …

Labilization And Diversification Of Pyrogenic Dissolved Organic Matter By Microbes, Aleksandar I. Goranov, Andrew S. Wozniak, Kyle W. Bostick, Andrew R. Zimmerman, Siddhartha Mitra, Patrick G. Hatcher

Chemistry & Biochemistry Faculty Publications

With the increased occurrence of forest fires around the world, interest in the chemistry of pyrogenic organic matter (pyOM) and its fate in the environment has increased. Upon leaching from soils by rain events, significant amounts of dissolved pyOM (pyDOM) enter the aquatic environment and interact with microbial communities that are essential for cycling organic matter within the different biogeochemical cycles. To evaluate the bio-reactivity of pyDOM, aqueous extracts of laboratory-produced chars were incubated with soil microbes and the molecular changes to the composition of pyDOM were probed using ultrahigh resolution mass spectrometry (Fourier transform – ion cyclotron resonance – …

Mass Spectral And Chemometric Analysis For The Detection And Identification Of Forensically Relevant Materials, Meghan Grace Appley

Legacy Theses & Dissertations (2009 - 2024)

Forensic science is the application of science to the law which in and of itself covers a substantial number of disciplines. Many of these involve the detection and identification of materials using analytical techniques. Traditional analysis methods include microscopy, spectroscopy and mass spectrometry. The materials encountered by forensic practitioners are ever evolving, resulting in the need to develop new methods to complement conventional techniques to enhance the evidentiary value of the samples. The research presented here shows that novel methods including direct analysis in real time–high-resolution mass spectrometry (DART-HRMS), thermal desorption coupled with gas chromatography-mass spectrometry (TD-GC-MS), solid-phase microextraction (SPME) …

Applications Of Mass Spectrometry To Forensic Entomology : Calliphoridae Species Determination, Monitoring Of Decomposition Volatiles, And Discovery Of Chemical Cues, Justine Elizabeth Giffen-Lemieux

Legacy Theses & Dissertations (2009 - 2024)

Forensic entomology is the study of insects that colonize human/animal wounds and remains and their development for use in legal contexts. It has numerous facets and applications including: estimation of time since death; investigation of neglect/abuse of children and the elderly; and determination of the presence of toxins and drugs as a potential cause of death. In order for the discovered insects to be of forensic utility, accurate species identification is crucial. Current methodologies for species determination rely on the knowledge and expertise of an entomologist; exhaustive taxonomic keys; and access to adequate equipment and facilities; resources which are often …

Application Of Direct Analysis In Real Time-High Resolution Mass Spectrometry To Genus- And Species-Level Differentiation And Identification Of Coral, Jessica Mary Hayes

Legacy Theses & Dissertations (2009 - 2024)

Coral reefs are one of the most biologically diverse ecosystems, and the accurate identification of the species is essential for diversity assessment, species delimitation and conservation. Current species and genus determination approaches are time consuming, resource intensive and can be highly subjective. To explore the hypothesis that the small molecule profiles of coral are species and genus specific, and can be used as a rapid tool to catalogue, classify and identify coral genera and species, the small-molecule chemical fingerprints of coral belonging to the species Acanthastrea echinata, Catalaphyllia jardinei, Duncanopsammia axifuga, Echinopora lamellosa, Euphyllia divisa, Euphyllia paraancora, Euphyllia paradivisa, Galaxea …

Novel Applications Of Mass Spectrometry For Quantitation And Reaction Mechanism Elucidation, Pengyi Zhao

Dissertations

Mass spectrometry (MS) has been growing as one of the most widely used tools in the field of analytical chemistry. Various applications have been developed to harness the high sensitivity and specificity of mass spectrometric analysis. In this dissertation, two major challenges are addressed. By developing mass spectrometric-based methods, absolute quantitation of proteins/peptides have been achieved. Elucidation of various reaction mechanisms are also enabled. These are the focuses of this dissertation.

In Chapters 2 to 4, a novel quantitation method is developed, titled as coulometric mass spectrometry (CMS). The strength of this method is that no reference standard or isotope-labeled …

Effects Of Oxidative Modifications On The Structure And Non-Canonical Functions Of Cytochrome C Studied By Mass Spectrometry, Victor Yin

Electronic Thesis and Dissertation Repository

The peroxidase activity of the mitochondrial protein cytochrome c (cyt c) plays a critical role in triggering programmed cell death, or apoptosis. However, the native structure of cyt c should render this activity impossible due to the lack of open iron coordination sites at its heme cofactor. Despite its key biological importance, the molecular mechanisms underlying this structure-function mismatch remain enigmatic. The work detailed in this dissertation fills this knowledge gap by using mass spectrometry (MS) to decipher the central role that protein oxidative modifications and their associated structural changes play in activating the peroxidase function of cyt c …

Analysis Of Secondary Metabolites Biosynthesized By Pathogenic And Symbiotic Fungi Using High-Resolution Tandem Lc-Ms And Spectral Molecular Networking, Natasha Desrochers

Electronic Thesis and Dissertation Repository

Fungi from Canadian crops affect many aspects of agriculture. For example, ginseng root rot caused by Ilyonectria spp. is a major issue for farmers. Ilyonectria spp. make few reported natural products and it is not known if those products are unique to virulent species. PCA and molecular networking were applied to HRMS data to establish a distinct metabolomic profile of root rot pathogens, characterized by antifungal resorcyclic acid lactones (RALs). These likely protect Ilyonectria from other soil pathogens, which gives it the opportunity to infect ginseng root.

Molecular networking was also applied to fungal endophytes from fruit crops to identify …

Structural Characterization Of Biomolecules Using Tandem Non- Linear And Linear Ion Mobility, Action Spectroscopy, Mass Spectrometry And Molecular Dynamics, Jacob Edward Porter

FIU Electronic Theses and Dissertations

The following thesis comprises the use of novel gas-phase spectroscopic and spectrometric techniques complemented with molecular dynamics for the structural characterization of biomolecules: from small model systems to large proteins. In particular, complementary linear and non-linear ion mobility, mass spectrometry and action spectroscopy has been applied to the separation and characterization of native and kinetic intermediate states of biomolecular ions. This suite of tools, comprising ion mobility in tandem with non-ergodic fragmentation and action spectroscopy, provided unique information on the conformational space, folding and binding dynamics of single stranded DNA, peptides, peptide-cofactor and protein systems. Theoretical modeling provided candidate structures …

From Neutral Aniline To Aniline Trication: A Computational And Experimental Study, G. L. Gutsev, H. A. López Peña, S. L. Mcpherson, Derrick Ampadu Boateng, B. R. Ramachandran, L. G. Gutsev, Katharine M. Tibbetts

Chemistry Publications

We report density functional theory computations and photoionization mass spectrometry measurements of aniline and its positively charged ions. The geometrical structures and properties of the neutral, singly, doubly, and triply positively charged aniline are computed using density functional theory with the generalized gradient approximation. At each charge, there are multiple isomers closely spaced in total energy. Whereas the lowest energy states of both neutral and cation have the same topology C₆H₅–NH₂, the dication and trication have the C₅NH₅–CH₂ topology with the nitrogen atom in the meta and para positions, …

Dissociation Of Singly And Multiply Charged Nitromethane Cations: Femtosecond Laser Mass Spectrometry And Theoretical Modeling, G. L. Gutsev, S. L. Mcpherson, Hugo A. López Peña, Derrick Ampadu Boateng, L. G. Gutsev, B. R. Ramachandran, Katharine M. Tibbetts

Chemistry Publications

Dissociation pathways of singly- and multiply charged gas-phase nitromethane cations were investigated with strong-field laser photoionization mass spectrometry and density functional theory computations. There are multiple isomers of the singly charged nitromethane radical cation, several of which can be accessed by rearrangement of the parent CH3–NO2 structure with low energy barriers. While direct cleavage of the C–N bond from the parent nitromethane cation produces NO2+ and CH3+, rearrangement prior to dissociation accounts for fragmentation products including NO+, CH2OH+, and CH2NO+. Extensive Coulomb explosion in fragment ions observed at high laser intensity indicates that rapid dissociation of multiply charged nitromethane cations …

Development And Implementation Of A Novel Voltage-Free Interface For Capillary Electrophoresis-Mass Spectrometry Separations Of Proteins, Courtney J. Kristoff

Graduate Theses, Dissertations, and Problem Reports

Capillary electrophoresis-mass spectrometry is a powerful technique for high-throughput and high efficiency separations combined with structural identification. Electrospray ionization is the primary interface used to couple capillary electrophoresis to mass analyzers; however, improved designs continue to be reported. A new interfacing method based on vibrating sharp-edge spray ionization is presented in this work to overcome the challenges of decoupling applied voltages and to enhance the compatibility with separations performed at near-neutral pH. The versatility and ease of use of this ionization source is demonstrated using β-blockers, peptides, and proteins. The cationic β-blocker pindolol was injected electrokinetically and detected at concentrations …

Genetic And Biochemical Characterization Of Ergot Alkaloid Synthesizing Fungi And Their Symbionts, Matthew Duane Maust

Graduate Theses, Dissertations, and Problem Reports

Ergot alkaloids are fungal tryptophan derived toxins which affect mammalian circulation and neurotransmission. These compounds are biosynthesized by a conserved genetic pathway, known as the ergot alkaloid synthesis (EAS) pathway by fungi belonging to the ascomycete families Trichocomaceae and Clavicipitaceae. Several Ipomoea species and related plants in the morning glory family harbor vertically transmitted symbiotic fungi in the genus Periglandula, also members of Clavicipitaceae, that produce ergot alkaloids. Metabolomic analysis of seeds identified a previously uncharacterized glycoside form of the pharmaceutically important ergot alkaloid, ergonovine. Several species belonging to the fungal genus Metarhizium have recently been shown to have the …

Protein/Peptide Characterization Using Mass Spectrometry And Molecular Dynamics Simulations, Ahmad Kiani Karanji

Graduate Theses, Dissertations, and Problem Reports

Mass spectrometry (MS) based-techniques and molecular dynamics (MD) simulations have been used to characterize protein/peptide structure as well as their interactions with lipid vesicles and detergents. Chapter 1 introduces an introduction to the concepts and tools that were used in this work. In Chapter 2, the dominant gas-phase conformer of [M+3H]3+ ions of the model peptide Acetyl-PSSSSKSSSSKSSSSKSSSSK are examined with ion mobility spectrometry (IMS), gas-phase hydrogen deuterium exchange (HDX), and mass spectrometry (MS) techniques. This section furthers the development of a protein structural prediction tool by providing information about gas-phase ion conformers of two model peptides having different solution conformational …

Protein Detection And Structural Characterization By Mass Spectrometry Using Supramolecular Assemblies And Small Molecules, Bo Zhao

Doctoral Dissertations

Mass spectrometry (MS) has played an increasingly prominent role in proteomics and structure biology because it shows superior capabilities in identification, quantification and structural characterization of proteins. To realize its full potential in protein analysis, significant progress has been made in developing innovative techniques and reagents that can couple to MS detection. This dissertation demonstrates the use of polymeric supramolecular assemblies for enhanced protein detection in complex biological mixtures by MS. An amphiphilic random co-polymer scaffold is developed to form functional supramolecular assemblies for protein/ peptide enrichment. The influences of charge density and functional group pK_a on host-guest interactions …

Molecular Recognition Of Methionine-Terminated Peptides By Cucurbit[8]Uril, Zoheb Hirani, Hailey F. Taylor, E. F. Babcock, Andrew T. Bockus, C. D. Varnado Jr., Christopher W. Bielawski, Adam R. Urbach

Adam R Urbach

This Article describes the molecular recognition of peptides containing an N-terminal methionine (Met) by the synthetic receptor cucurbit[8]-uril (Q8) in aqueous solution and with submicromolar affinity. Prior work established that Q8 binds with high affinity to peptides containing aromatic amino acids, either by simultaneous binding of two aromatic residues, one from each of two different peptides, or by simultaneous binding of an aromatic residue and its immediate neighbor on the same peptide. The additional binding interface of two neighboring residues suggested the possibility of targeting nonaromatic peptides, which have thus far bound only weakly to synthetic receptors. A peptide library …

Development Of A Screening Method For Drugs Of Abuse By Direct Analysis Of Dried Urine Spots Coupled To Mass Spectrometry, Melanie Goldstein

Student Theses

Fast and easy screening procedures are essential in any forensic toxicology laboratory to differentiate negative samples from presumptive positive cases. Immunoassay techniques, such as enzyme multiplied immunoassay technique (EMIT), are routinely employed as screening procedures. However, these techniques lack specificity (only differentiate group of drugs and not individual compounds) and it is difficult to add new compounds to the panel. Direct analysis of dried urine spots (DUS) by mass spectrometry (MS) offers a novel strategy to overcome these issues. DUS offer an improved storage alternative for biological samples, reducing costs and space requirements. In this work, an original method to …

Lipid Biomarkers Of Bering And Chukchi Sea Euphausiids And Their Application To Diet History, Rachel L. Pleuthner

OES Theses and Dissertations

In the eastern Bering Sea, Thysanoessa raschii are the most abundant krill species and a keystone trophic member that serve as both an important grazer and link to upper level consumers. In this system krill experience large annual variation in food resources, especially during ice advance and retreat; multiple lipid classes are used to temper the effects of those fluctuations, as well as to fuel reproduction and growth. Two shipboard feeding experiments that occurred during late spring and early summer of 2010, respectively, monitored the lipid retention in adult T. raschii and examined the fluctuation of specific lipid biomarkers under …

Nitrogen Heterocycles Form Peptide Nucleic Acid Precursors In Complex Prebiotic Mixtures, Laura E. Rodriguez, Christopher H. House, Karen E. Smith, Melissa R. Roberts, Michael P. Callahan

Chemistry and Biochemistry Faculty Publications and Presentations

The ability to store information is believed to have been crucial for the origin and evolution of life; however, little is known about the genetic polymers relevant to abiogenesis. Nitrogen heterocycles (N-heterocycles) are plausible components of such polymers as they may have been readily available on early Earth and are the means by which the extant genetic macromolecules RNA and DNA store information. Here, we report the reactivity of numerous N-heterocycles in highly complex mixtures, which were generated using a Miller-Urey spark discharge apparatus with either a reducing or neutral atmosphere, to investigate how N-heterocycles are modified under plausible prebiotic …